Data supporting "PHEIGES, all-cell-free phage synthesis and selection from engineered genomes"

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2022-08-30
2023-12-01

Date completed

2024-01-17

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Data supporting "PHEIGES, all-cell-free phage synthesis and selection from engineered genomes"

Published Date

2024-01-29

Author Contact

Noireaux, Vincent
noireaux@umn.edu

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Dataset
Experimental Data
Genomics Data

Abstract

Bacteriophages constitute an invaluable biological reservoir for biotechnology and medicine. The ability to exploit such vast resources is hampered by the lack of methods to rapidly engineer, assemble, package genomes, and select phages. Cell-free transcription-translation (TXTL) offers experimental settings to address such a limitation. Here, we describe PHage Engineering by In vitro Gene Expression and Selection (PHEIGES) using T7 phage genome and Escherichia coli TXTL. Phage genomes are assembled in vitro from PCR-amplified fragments and directly expressed in batch TXTL reactions to produce up to 1011 PFU/ml engineered phages within one day. We further demonstrate a significant genotype-phenotype linkage of phage assembly in bulk TXTL. This enables rapid selection of phages with altered rough lipopolysaccharides specificity from phage genomes incorporating tail fiber mutant libraries. We establish the scalability of PHEIGES by one pot assembly of such mutants with fluorescent gene integration and 10% length-reduced genome.

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Microsoft Excel files with experimental data (microscopy data) DNA sequencing data (NGS sequencing)

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Funding information

National Science Foundation CBET FMRG 2228971
Fulbright U.S. Student Program, which is sponsored by the U.S. Department of State, the Romanian-U.S. Fulbright Commission, and the Franco-American Fulbright Commission
Bettencourt Schueller Foundation

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Previously Published Citation

Suggested citation

Levrier, Antoine; Karpathakis, Ioannis; Nash, Bruce; Bowden, Steven; Lindner, Ariel; Noireaux, Vincent. (2024). Data supporting "PHEIGES, all-cell-free phage synthesis and selection from engineered genomes". Retrieved from the Data Repository for the University of Minnesota (DRUM), https://doi.org/10.13020/0CDV-Q695.
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File View/OpenDescriptionSize
movie raw images.zipSupplementary Movie: Raw microscopy acquisitions for E. coli B infection by T7-mC-gp10 timelapse.448.21 MB
compil raw data.zipFigure 6.c : Supplementary Data micrographs for E. coli rfaC infected by T7-compile24.13 MB
NGS_illumina.zipFigure 5.c : Raw NGS data for mutagenic PCR tip of the tail fiber fragments.1.93 GB
raw-sequencing.zipAll the raw sequencing obtained for the 26 fully sequenced phages and the alignment files for each phage genome.13.79 MB
Readme_Noireaux_2024.txtDescription of the data7.46 KB

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